Day 3: Difference between revisions

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New page: 2. Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in...
 
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2.
2.Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in the presence of kanamycin, meaning the DNA was not taken up by the cell.
 
Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in the presence of kanamycin, meaning the DNA was not taken up by the cell.


Outcome #2, thousands of colonies on all plates: This could indicate contamination because the cells transformed with the negative control for ligation, backbone only and no ligase, should not have kanamycin resistance.
Outcome #2, thousands of colonies on all plates: This could indicate contamination because the cells transformed with the negative control for ligation, backbone only and no ligase, should not have kanamycin resistance.


Outcome #3, approximately the same number of colonies on the backbone+ligase+kill cut as the backbone+insert+ligase+kill cut: This could also indicate that there wasn't complete digestion during the kill-cut and some backbone managed to back-ligate and be transformed into the cells of both samples.
Outcome #3, approximately the same number of colonies on the backbone+ligase+kill cut as the backbone+insert+ligase+kill cut: This could also indicate that there wasn't complete digestion during the kill-cut and some backbone managed to back-ligate and be transformed into the cells of both samples.
3.
{| border="1"
! Diagnostic digest 1
| plasmid with insert
| plasmid no insert
|-
| Enzyme(s) used
|
|
|-
| Buffer used
|
|
|-
| Temperature
|
|
|-
| Predicted fragments
|
|
|-
|
|
|
|-
! Diagnostic digest 2
|
|
|-
| Enzyme(s) used
|
|
|-
| Buffer used
|
|
|-
| Temperature
|
|
|-
| Predicted fragments
|
|
|}

Revision as of 20:34, 20 September 2007

2.Outcome #1, no colonies on any plate: This would suggest that the transformation was unsuccessful, because even the positive control with the un-engineered M13K07 DNA failed to grow in the presence of kanamycin, meaning the DNA was not taken up by the cell.

Outcome #2, thousands of colonies on all plates: This could indicate contamination because the cells transformed with the negative control for ligation, backbone only and no ligase, should not have kanamycin resistance.

Outcome #3, approximately the same number of colonies on the backbone+ligase+kill cut as the backbone+insert+ligase+kill cut: This could also indicate that there wasn't complete digestion during the kill-cut and some backbone managed to back-ligate and be transformed into the cells of both samples.

3.

Diagnostic digest 1 plasmid with insert plasmid no insert
Enzyme(s) used
Buffer used
Temperature
Predicted fragments
Diagnostic digest 2
Enzyme(s) used
Buffer used
Temperature
Predicted fragments